Extended Data Fig. 2: Generation of Tgnd6 iKD and Tgnd9 iKD.
From: An Alveolata secretory machinery adapted to parasite host cell invasion
a, Strategy for conditional depletion of TgNd6 (TGGT1_248640) using the Auxin-inducible degron system. b, Integration PCRs showing double homologous recombination at the 5’ (top) and at the 3’ (bottom) of the Tgnd6 locus. Representative of 3 experiments. c, Strategy for conditional depletion of TgNd9 (TGGT1_249730) using the Tet-OFF system. d, Integration PCRs showing double homologous recombination at the 3’ (top) and at the 5’ (bottom) of the Tgnd9 locus. Representative of 3 experiments. e, Immunoblot of Tgnd6 iKD ± IAA (top) and Tgnd9 iKD ± ATc (bottom) showing the timing for protein depletion. ROP5, loading control. Ctrl: ΔKu80-Tir1 (upper panel) and ΔKu80-TaTi (lower panel). Representative of one experiment for Tgnd9 iKD and 2 experiments for Tgnd6 iKD. f, IFA of Tgnd6 iKD shows depletion of TgNd6 after 24 hours IAA treatment (top right), IFA of Tgnd9 iKD shows depletion of TgNd9 after 72 hours ATc treatment (bottom right). Red, anti-ARO. Green, anti-HA. Representative of 3 experiments. g, Detergent extraction profile of TgND6 and TgND9 after solubilization of HA‐tagged TgNds parasites with Tris 1 mM, 1% TritonX‐100 or 1% SDS. Representative of 2 experiments.
